A capillary electrophoresis laser-induced fluorescence method for analysis of potato glycoalkaloids based on a solution-phase immunoassay. 1. Separation and quantification of immunoassay products |
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Authors: | Driedger D R LeBlanc R J LeBlanc E L Sporns P |
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Institution: | Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, AB, T6G 2P5 Canada. |
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Abstract: | Solution-phase immunoassays are typically faster and more precise than ELISAs. This research developed a solution-phase for the immunoassay of potato glycoalkaloids (GAs) based on quantification by capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection. Solanidine coupled to 4'-(aminomethyl)fluorescein and a polyclonal antibody solution were used as the immunoreagents. Unbound fluorescent solanidine was detected by CE-LIF (excitation 488 nm, emission 520 nm). Optimum resolution of immunoassay products was achieved with a buffer consisting of 50 mM phosphate, 10% (v/v) methanol, and 1.5 mM SDS, pH 7.5. A plot of signal vs log GA] produced a sigmoidal curve typical of immunoassays. Analysis of extracts of sprouted Yukon Gold potato tubers and nonsprouted Yukon Gold tubers resulted in total GA] of 98 microg/g (RSD 9%) and 55 microg/g (RSD 9%), respectively. The findings indicated that CE-LIF coupled with a solution-phase immunoassay can be used to quantify total GA in potatoes. |
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